Studying the Heat Effect

of Nanoparticles

Dr. Jesús M. de la Fuente

Instituto de Ciencia de Materiales de Aragon,

CSIC/University of Zaragoza, Spain

e-mail: jmfuente@unizar.es

IN VIVO MAGNETIC HYPERTHERMIA DEVICE

IN VIVO MAGNETIC HYPERTHERMIA GENERAL PROCEDURE

MNPsIntratumoral injecction

2,5 105 EL4 cellsSubcutaneous

7 days

AC exposure 30min105 kHz, 225 GaussDays 0, 1, 2 post-MNPs inyection

Sacrifice5 days-post-MNPsInyectionTumour and blood extraction

Daily tumour volume measurementGroups (5 mice/each):- Control- AC control- MNPs control- AC+MNPs

Nuclei were stained with DAPI. Scale bar: 50 µm.

PREVIOUS RESULTS WITH OUR MNPs (80ugFe/mouse)

A B

C,D E,F

Tumour volumes are normalized to the tumour volume on day 1when the injection was made. Error bars represent SEM. (*p < 0.05;two-way ANOVA, Tukey’s multiple comparisons test.

EL4

107

Subcutaneou

sIntratumora

l0

2,5x105 cell/

100 μL

C57BL/6

1mg material/50 μL sterile PBS

(95,5 µgFe/mouse)

days

Sacrifice

Tumour and

blood extraction

Histopathological

analysis

Perls, HE, DAPI

MNPs (EG5115Rho@CPS)

3 mice controls

3 mice MNPs

EXPERIMENTAL PROCEDURE TO ANALYZE MNPs DISTRIBUTION IN THE TUMOUR

MNPs DISTRIBUTION IN THE TUMOUR- FLUORESCENCE

Nuclei-DAPIMNPs-RHO

Results Perls (Fe3+ staining)

MNPs DISTRIBUTION IN THE TUMOUR- PERLS

3D reconstruction

MNPs DISTRIBUTION IN THE TUMOUR- CONFOCAL FLUORESCENCE

MNPs were detecteddeeply in the tumour mass

MNPs DISTRIBUTION IN THE TUMOUR- CONFOCAL FLUORESCENCE

IN VIVO MAGNETIC HYPERTHERMIA: EXPERIMENT IN PROGRESS

MNPsIntratumoral injecction

2,5 105 EL4 cellsSubcutaneous

7 days

AC exposure 30min105 kHz, 225 GaussDays 0, 1, 2 post-MNPs inyection

Sacrifice5 days-post-MNPsInyectionTumour and blood extraction

Daily tumour volume measurement

MNPs-Doxo (EG5115Rho@CPS DOXO)

CONTROL MNPs- DOXO MNPs DOXO

✅Done in previousexperiments

IN PROGRESSFUTURE EXPERIMENTS

5 MICE/GROUP

PREPARACIÓN DE LAS MNPS

-EG5115Rho@CPS (Ref interna LA_NP_EG_1)-Se piensa inyectar 1mg material/ratón-Pesamos 10mg de material-Resuspendemos en 1mL de EtOh 70% con vortex y sonicación-Lavamos 5 veces con PBS estéril. Centrifugando 5min 14000rpm-En el último lavado resuspendo en 500 µL (20 mg material/mL)-Se preparan las MNPs el mismo día de la inyección, cuando se van a pinchar presentan buena dispersión-Guardo la suspensión de MNPs sobrante y analizamos contenido en hierro:

0,35 mgFe/mL1,91

-Inyectando 50 µL/ratón = 95,5 µgFe/ratón-Las MNPs tienen una relación: 95,5 µgFe/mg material